title: The structure of the neisserial lipooligosaccharide phosphoethanolamine transferase A (LptA) required for resistance to polymyxin.
abstract: Gram-negative bacteria possess an outer membrane envelope consisting of an outer leaflet of lipopolysaccharides, also called endotoxins, which protect the pathogen from antimicrobial peptides and have multifaceted roles in virulence. Lipopolysaccharide consists of a glycan moiety attached to lipid A, embedded in the outer membrane. Modification of the lipid A headgroups by phosphoethanolamine (PEA) or 4-amino-arabinose residues increases resistance to the cationic cyclic polypeptide antibiotic, polymyxin. Lipid A PEA transferases are members of the YhjW/YjdB/YijP superfamily and usually consist of a transmembrane domain anchoring the enzyme to the periplasmic face of the cytoplasmic membrane attached to a soluble catalytic domain. The crystal structure of the soluble domain of the protein of the lipid A PEA transferase from Neisseria meningitidis has been determined crystallographically and refined to 1.4Å resolution. The structure reveals a core hydrolase fold similar to that of alkaline phosphatase. Loop regions in the structure differ, presumably to enable interaction with the membrane-localized substrates and to provide substrate specificity. A phosphorylated form of the putative nucleophile, Thr280, is observed. Metal ions present in the active site are coordinated to Thr280 and to residues conserved among the family of transferases. The structure reveals the protein components needed for the transferase chemistry; however, substrate-binding regions are not evident and are likely to reside in the transmembrane domain of the protein.
authors.affiliation: School of Chemistry and Biochemistry, University of Western Australia, 35 Stirling Highway, Crawley, WA 6009, Australia.
authors.forename: Christopher
authors.initials: C
authors.lastname: Wanty
authors.forename: Anandhi
authors.initials: A
authors.lastname: Anandan
authors.forename: Susannah
authors.initials: S
authors.lastname: Piek
authors.forename: James
authors.initials: J
authors.lastname: Walshe
authors.forename: Jhuma
authors.initials: J
authors.lastname: Ganguly
authors.forename: Russell W
authors.initials: RW
authors.lastname: Carlson
authors.forename: Keith A
authors.initials: KA
authors.lastname: Stubbs
authors.forename: Charlene M
authors.initials: CM
authors.lastname: Kahler
authors.forename: Alice
authors.initials: A
authors.lastname: Vrielink
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001665:Binding Sites; D024881:Drug Resistance, Bacterial; D004982:Ethanolaminephosphotransferase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004983:Ethanolamines / Q000378:metabolism; D008050:Lipid A / Q000378:metabolism; D008070:Lipopolysaccharides / Q000378:metabolism; D008954:Models, Biological; D008958:Models, Molecular; D009345:Neisseria meningitidis / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D011113:Polymyxins / Q000494:pharmacology*; D054730:Protein Interaction Domains and Motifs / Q000235:genetics; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary / Q000502:physiology
chemical_list: D000900:Anti-Bacterial Agents; D004983:Ethanolamines; D008050:Lipid A; D008070:Lipopolysaccharides; D011113:Polymyxins; C023023:lipid-linked oligosaccharides; C005448:phosphorylethanolamine; D004982:Ethanolaminephosphotransferase
keywords: AlkPP; EDTA; LPS; LptA; MAD; MALDI-TOF-MS; PDB; PEA; PEG; Protein Data Bank; SeMet; alkaline phosphatase; crystallography; endotoxin biosynthesis; ethylenediaminetetraacetic acid; lipid A phosphoethanolamine transferase; lipopolysaccharide; matrix-assisted laser/desorption time-of-flight mass spectrometry; multiple-wavelength anomalous diffraction; p-NPPE; para-nitrophenyl phosphoethanolamine; phosphoethanolamine; phosphoethanolamine transferase; polyethylene glycol; polymyxin resistance; selenomethionine